CRISPR/Cas9 Technology for Potato Functional Genomics and Breeding.

Cultivated potato (Solanum tuberosum L.) is one of the most important staple food crops worldwide. Its tetraploid and highly heterozygous nature poses a great challenge to its basic research and trait improvement through traditional mutagenesis and/or crossbreeding. The establishment of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) as a gene editing tool has allowed the alteration of specific gene sequences and their concomitant gene function, providing powerful technology for potato gene functional analysis and improvement of elite cultivars. This technology relies on a short RNA molecule called single guide RNA (sgRNA) that directs the Cas9 nuclease to induce a site-specific double-stranded break (DSB). Further, repair of the DSB by the error-prone non-homologous end joining (NHEJ) mechanism leads to the introduction of targeted mutations, which can be used to produce the loss of function of specific gene(s). In this chapter, we describe experimental procedures to apply the CRISPR/Cas9 technology for potato genome editing. First, we provide strategies for target selection and sgRNA design and describe a Golden Gate-based cloning system to obtain a sgRNA/Cas9-encoding binary vector. We also describe an optimized protocol for ribonucleoprotein (RNP) complex assembly. The binary vector can be used for both Agrobacterium-mediated transformation and transient expression in potato protoplasts, while the RNP complexes are intended to obtain edited potato lines through protoplast transfection and plant regeneration. Finally, we describe procedures to identify the gene-edited potato lines. The methods described here are suitable for potato gene functional analysis and breeding.

[Delirium, catatonia and substance withdrawal syndrome manifested as psychomotor agitation in COVID-19: a pharmacological management approach for the general hospital setting]. / Delirium, catatonía y síndrome de abstinencia manifestados como agitación psicomotora en COVID-19: una propuesta de manejo farmacológico en el hospital general.

BACKGROUND: Neuropsychiatric symptoms can be part of the clinical spectrum of COVID-19 infections. AIM: To devise an evidence based clinical algorithm as a guide for clinicians, to identify and treat underlying clinical syndromes of psychomotor agitation, such as delirium, catatonia or substance withdrawal in patients who are hospitalized and infected with SARS-CoV-2. MATERIAL AND METHODS: A review of the literature about the pharmacological management of neuropsychiatric manifestations of COVID-19 at the general hospital, to develop a clinical protocol based on a consensus from an interdisciplinary expert panel at a Clinical Hospital. RESULTS: A consensual clinical algorithm for the management of delirium, catatonia, and substance withdrawal, manifested as psychomotor agitation in patients hospitalized with COVID-19, was developed as a clinical proposal for physicians at different levels of complexity in health services. CONCLUSIONS: Cooperation among different clinical units in the general hospital facilitated the implementation of a clinical algorithm for clinicians for the management of psychomotor agitation in COVID-19 patients.

Delirium, catatonía y síndrome de abstinencia manifestados como agitación psicomotora en COVID-19: una propuesta de manejo farmacológico en el hospital general / Delirium, catatonia and substance withdrawal syndrome manifested as psychomotor agitation in COVID-19: a pharmacological management approach for the general hospital setting

BACKGROUND: Neuropsychiatric symptoms can be part of the clinical spectrum of COVID-19 infections. AIM: To devise an evidence based clinical algorithm as a guide for clinicians, to identify and treat underlying clinical syndromes of psychomotor agitation, such as delirium, catatonia or substance withdrawal in patients who are hospitalized and infected with SARS-CoV-2. MATERIAL AND METHODS: A review of the literature about the pharmacological management of neuropsychiatric manifestations of COVID-19 at the general hospital, to develop a clinical protocol based on a consensus from an interdisciplinary expert panel at a Clinical Hospital. Results: A consensual clinical algorithm for the management of delirium, catatonia, and substance withdrawal, manifested as psychomotor agitation in patients hospitalized with COVID-19, was developed as a clinical proposal for physicians at different levels of complexity in health services. CONCLUSIONS: Cooperation among different clinical units in the general hospital facilitated the implementation of a clinical algorithm for clinicians for the management of psychomotor agitation in COVID-19 patients.

Comparación de la capacidad de salto en deportistas juveniles / Comparação da capacidade de salto em atletas juvenis / Comparison about the Jump capacity in junior athletes

RESUMEN Dentro de la dinámica del entrenamiento deportivo la evaluación de salto es una herramienta útil para determinar y comparar el rendimiento del atleta. El objetivo del estudio es comparar la capacidad de salto entre dos grupos de deportistas juveniles, que ingresan a la universidad en diferente año. El estudio es de tipo comparativo, se seleccionaron 120 sujetos por conveniencia, los cuales fueron distribuidos en dos grupos, de acuerdo con el año en el que ingresan a la universidad (2015 y 2020). Se midió capacidad de salto con los protocolos de sentadilla de 90°, contra movimiento y libre. Los resultados mostraron una tendencia (p=0,07) en la modalidad de contra movimiento, donde el grupo del año 2020 poseía un mejor rendimiento. En conclusión, el salto en contra movimiento se proyecta como la variable de comparación para poblaciones de similares características.

RESUMO Dentro da dinâmica do treinamento esportivo, a avaliação do salto é uma ferramenta útil para determinar e comparar o desempenho do atleta. O objetivo do estudo é comparar a capacidade de salto entre dois grupos de jovens atletas, que iniciam a universidade em anos diferentes. O estudo é de tipo comparativo, 120 sujeitos foram selecionados por conveniência, os quais se distribuíram em dois grupos, de acordo com o ano em que ingressaram na universidade (2015 e 2020). A capacidade de salto foi medida com os protocolos de 90° de agachamento, contra-movimento e agachamento livre. Os resultados mostraram uma tendência (p=0,07) na modalidade de contra-movimento, onde o grupo de 2020 teve um melhor desempenho. Em conclusão, o salto de contra-movimento é projetado como a variável de comparação para populações com características semelhantes.

ABSTRACT Within the dynamics of sports training the jumping evaluation is a useful tool to determine and compare the athlete's performance. The objective of the study is to compare the jumping ability between two groups of juvenile athletes, entering university in different years. The study is of a comparative type, 120 subjects were selected by convenience, which was distributed in two groups, according to the year in which they enter the university (2015 and 2020). Jumping capacity was measured with the 90° squat, countermovement and free squat protocols. The results showed a trend (p=0.07) in the countermovement modality, where the 2020 year group possessed better performance. In conclusion, the countermovement jump is projected as the comparison variable for populations with similar characteristics.

Molecular Detection of Ralstonia solanacearum to Facilitate Breeding for Resistance to Bacterial Wilt in Potato.

Potato bacterial wilt is caused by the devastating bacterial pathogen Ralstonia solanacearum. Quantitative resistance to this disease has been and is currently introgressed from a number of wild relatives into cultivated varieties through laborious breeding programs. Here, we present two methods that we have developed to facilitate the screening for resistance to bacterial wilt in potato. The first one uses R. solanacearum reporter strains constitutively expressing the luxCDABE operon or the green fluorescent protein (gfp) to follow pathogen colonization in potato germplasm. Luminescent strains are used for nondestructive live imaging, while fluorescent ones enable precise pathogen visualization inside the plant tissues through confocal microscopy. The second method is a BIO-multiplex-PCR assay that is useful for sensitive and specific detection of viable R. solanacearum (IIB-1) cells in latently infected potato plants. This BIO-multiplex-PCR assay can specifically detect IIB-1 sequevar strains as well as strains belonging to all four R. solanacearum phylotypes and is sensitive enough to detect without DNA extraction ten bacterial cells per mL in complex samples.The described methods allow the detection of latent infections in roots and stems of asymptomatic plants and were shown to be efficient tools to assist potato breeding programs.

State of the Art of Genetic Engineering in Potato: From the First Report to Its Future Potential.

Potato (Solanum tuberosum L.) is a crop of world importance that produces tubers of high nutritional quality. It is considered one of the promising crops to overcome the challenges of poverty and hunger worldwide. However, it is exposed to different biotic and abiotic stresses that can cause significant losses in production. Thus, potato is a candidate of special relevance for improvements through conventional breeding and biotechnology. Since conventional breeding is time-consuming and challenging, genetic engineering provides the opportunity to introduce/switch-off genes of interest without altering the allelic combination that characterize successful commercial cultivars or to induce targeted sequence modifications by New Breeding Techniques. There is a variety of methods for potato improvement via genetic transformation. Most of them incorporate genes of interest into the nuclear genome; nevertheless, the development of plastid transformation protocols broadened the available approaches for potato breeding. Although all methods have their advantages and disadvantages, Agrobacterium-mediated transformation is the most used approach. Alternative methods such as particle bombardment, protoplast transfection with polyethylene glycol and microinjection are also effective. Independently of the DNA delivery approach, critical steps for a successful transformation are a rapid and efficient regeneration protocol and a selection system. Several critical factors affect the transformation efficiency: vector type, insert size, Agrobacterium strain, explant type, composition of the subculture media, selective agent, among others. Moreover, transient or stable transformation, constitutive or inducible promoters, antibiotic/herbicide resistance or marker-free strategies can be considered. Although great efforts have been made to optimize all the parameters, potato transformation protocols are highly genotype-dependent. Genome editing technologies provide promising tools in genetic engineering allowing precise modification of targeted sequences. Interestingly, transient expression of genome editing components in potato protoplasts was reported to generate edited plants without the integration of any foreign DNA, which is a valuable aspect from both a scientific and a regulatory perspective. In this review, current challenges and opportunities concerning potato genetic engineering strategies developed to date are discussed. We describe their critical parameters and constrains, and the potential application of the available tools for functional analyses or biotechnological purposes. Public concerns and safety issues are also addressed.

Neurological soft signs (NSS) and cognitive deficits in HIV associated neurocognitive disorder.

Neurological soft signs (NSS) are frequently found in severe mental disorders, such as Alzheimer's disease, schizophrenia or HIV associated neurocognitive disorder (HAND) which includes asymptomatic neurocognitive impairment (ANI), mild neurocognitive disorder (MND) and HIV-associated dementia. To characterize NSS in patients with HIV we examined them with respect to neuropsychological deficits typically found in the disorder. 67 HIV + patients without a history of head trauma, opportunistic infections, severe psychiatric disorders or acute confounding comorbidities of the Central nervous system (CNS) were recruited. NSS and neuropsychological deficits were examined on the Heidelberg scale and the Cambridge Neuropsychological Test Automated Battery (CANTAB), respectively. Semantic and phonemic verbal fluency were additionally established. According to NIMH and NINDS criteria, 18 patients were diagnosed with ANI and 21 with MND, 28 showed no cognitive deficits. NSS total scores were significantly correlated with several cognitive domains and NSS subscales. These correlations were confirmed when motor performance was entered as a covariate. According to our findings, NSS in HIV positive patients are significantly correlated with deficits in a broad range of neuropsychological domains. Similar findings were reported in schizophrenia, emphasizing the transdiagnostic character of NSS and supporting NSS examination in screening HIV patients for HAND.

Ephrin-B1 Is a Novel Biomarker of Bladder Cancer Aggressiveness. Studies in Murine Models and in Human Samples.

Bladder cancer (BC) is the ninth most common cancer worldwide, but molecular changes are still under study. During tumor progression, Epithelial cadherin (E-cadherin) expression is altered and ß-catenin may be translocated to the nucleus, where it acts as co-transcription factor of tumor invasion associated genes. This investigation further characterizes E-cadherin and ß-catenin associated changes in BC, by combining bioinformatics, an experimental murine cell model (MB49/MB49-I) and human BC samples. In in silico studies, a DisGeNET (gene-disease associations database) analysis identified CDH1 (E-cadherin gene) as one with highest score among 130 BC related-genes. COSMIC mutation analysis revealed CDH1 low mutations rates. Compared to MB49 control BC cells, MB49-I invasive cells showed decreased E-cadherin expression, E- to P-cadherin switch, higher ß-catenin nuclear signal and lower cytoplasmic p-Ser33-ß-catenin signal, higher Ephrin-B1 ligand and EphB2 receptor expression, higher Phospho-Stat3 and Urokinase-type Plasminogen Activator (UPA), and UPA receptor expression. MB49-I cells transfected with Ephrin-B1 siRNA showed lower migratory and invasive capacity than control cells (scramble siRNA). By immunohistochemistry, orthotopic MB49-I tumors had lower E-cadherin, increased nuclear ß-catenin, lower pSer33-ß-catenin cytoplasmic signal, and higher Ephrin-B1 expression than MB49 tumors. Similar changes were found in human BC tumors, and 83% of infiltrating tumors depicted a high Ephrin-B1 stain. An association between higher Ephrin-B1 expression and higher stage and tumor grade was found. No association was found between abnormal E-cadherin signal, Ephrin-B1 expression or clinical-pathological parameter. This study thoroughly analyzed E-cadherin and associated changes in BC, and reports Ephrin-B1 as a new marker of tumor aggressiveness.

Optimization of culture conditions for the expression of three different insoluble proteins in Escherichia coli.

Recombinant protein expression for structural and therapeutic applications requires the use of systems with high expression yields. Escherichia coli is considered the workhorse for this purpose, given its fast growth rate and feasible manipulation. However, bacterial inclusion body formation remains a challenge for further protein purification. We analyzed and optimized the expression conditions for three different proteins: an anti-MICA scFv, MICA, and p19 subunit of IL-23. We used a response surface methodology based on a three-level Box-Behnken design, which included three factors: post-induction temperature, post-induction time and IPTG concentration. Comparing this information with soluble protein data in a principal component analysis revealed that insoluble and soluble proteins have different optimal conditions for post-induction temperature, post-induction time, IPTG concentration and in amino acid sequence features. Finally, we optimized the refolding conditions of the least expressed protein, anti-MICA scFv, using a fast dilution protocol with different additives, obtaining soluble and active scFv for binding assays. These results allowed us to obtain higher yields of proteins expressed in inclusion bodies. Further studies using the system proposed in this study may lead to the identification of optimal environmental factors for a given protein sequence, favoring the acceleration of bioprocess development and structural studies.

Development of a new promoter to avoid the silencing of genes in the production of recombinant antibodies in chinese hamster ovary cells.

BACKGROUND: The production of recombinant proteins in mammalian cell lines is one of the most important areas in biopharmaceutical industry. Viral transcriptional promoters are widely used to express recombinant proteins in mammalian cell lines. However, these promoters are susceptible to silencing, thus limiting protein productivity. Some CpG islands can avoid the silencing of housekeeping genes; for that reason, they have been used to increase the production of recombinant genes in cells of animal origin. In this study, we evaluated the CpG island of the promoter region of the ß-actin gene of Cricetulus griseous (Chinese hamster), associated to the Cytomegalovirus (CMV) promoter, to increase recombinant antibodies production in Chinese Hamster Ovary (CHO) cells. RESULTS: We focused on the non-coding region of CpG island, which we called RegCG. RegCG behaved as a promoter, whose transcriptional activity was mainly commanded by the CAAT and CArG boxes of the proximal promoter. However, the transcription started mainly at the intronic region before the proximal transcription start site. While the CMV promoter was initially more powerful than RegCG, the latter promoter was more resistant to silencing than the CMV promoter in stable cell lines, and its activity was improved when combined with the CMV promoter. Thereby, the chimeric promoter was able to maintain the expression of recombinant antibodies in stable clones for 40 days at an average level 4 times higher than the CMV promoter. Finally, the chimeric promoter showed compatibility with a genetic amplification system by induction with methotrexate in cells deficient in the dihydrofolate reductase gene. CONCLUSIONS: We have generated an efficient synthetic hybrid transcription promoter through the combination of RegCG with CMV, which, in stable cell lines, shows greater activity than when both promoters are used separately. Our chimeric promoter is compatible with a genetic amplification system in CHO DG44 cells and makes possible the generation of stable cell lines with high production of recombinant antibodies. We propose that this promoter can be a good alternative for the generation of clones expressing high amount of recombinant proteins, essential for industrial applications.

Transcription factor engineering in CHO cells for recombinant protein production.

The continuous increase of approved biopharmaceutical products drives the development of more efficient recombinant protein expression systems. Chinese hamster ovary (CHO) cells are the mainstay for this purpose but have some drawbacks, such as low levels of expression. Several strategies have been applied to increase the productivity of CHO cells with different outcomes. Transcription factor (TF) engineering has emerged as an interesting and successful approach, as these proteins can act as master regulators; the expression and function of a TF can be controlled by small molecules, and it is possible to design tailored TFs and promoters with desired features. To date, the majority of studies have focused on the use of TFs with growth, metabolic, cell cycle or endoplasmic reticulum functions, although there is a trend to develop new, synthetic TFs. Moreover, new synthetic biological approaches are showing promising advances for the development of specific TFs, even with tailored ligand sensitivity. In this article, we summarize the strategies to increase recombinant protein expression by modulating and designing TFs and with advancements in synthetic biology. We also illustrate how this class of proteins can be used to develop more robust expression systems.

Reduced Enzymatic Browning in Potato Tubers by Specific Editing of a Polyphenol Oxidase Gene via Ribonucleoprotein Complexes Delivery of the CRISPR/Cas9 System.

Polyphenol Oxidases (PPOs) catalyze the conversion of phenolic substrates to quinones, leading to the formation of dark-colored precipitates in fruits and vegetables. This process, known as enzymatic browning, is the cause of undesirable changes in organoleptic properties and the loss of nutritional quality in plant-derived products. In potato (Solanum tubersoum L.), PPOs are encoded by a multi-gene family with different expression patterns. Here, we have studied the application of the CRISPR/Cas9 system to induce mutations in the StPPO2 gene in the tetraploid cultivar Desiree. We hypothesized that the specific editing of this target gene would result in a lower PPO activity in the tuber with the consequent reduction of the enzymatic browning. Ribonucleoprotein complexes (RNPs), formed by two sgRNAs and Cas9 nuclease, were transfected to potato protoplasts. Up to 68% of regenerated plants contained mutations in at least one allele of the target gene, while 24% of edited lines carried mutations in all four alleles. No off-target mutations were identified in other analyzed StPPO genes. Mutations induced in the four alleles of StPPO2 gene, led to lines with a reduction of up to 69% in tuber PPO activity and a reduction of 73% in enzymatic browning, compared to the control. Our results demonstrate that the CRISPR/Cas9 system can be applied to develop potato varieties with reduced enzymatic browning in tubers, by the specific editing of a single member of the StPPO gene family.

Functional and Objective Results of Urinary Undiversions in Oncologic Patients.

OBJECTIVE: To review surgical outcomes in urinary undiversions and describe short and long-term complications. METHODS: Retrospective review of patients who underwent urinary undiversion in our institution between May 2010 and May 2016. Complications were graded according to the Clavien classification. All patients completed the Patient Global Impression of Improvement questionnaire to indicate overall satisfaction with urinary undiversion. RESULTS: Median time from the cystectomy to undiversion was 29 months (range 11-53 months). Five patients (55%) reported significant distress related to the ileal conduit and were undiverted into an orthotopic neobladder. A female patient with an orthotopic neobladder and severe urinary incontinence received neobladder neck closure and catheterizable channel. Another female patient with an orthotopic neobladder was undiverted into an Indiana Pouch. Complications during the first 60 days were mostly minor, Clavien I (1 patient), 5 patients Clavien II, and a patient with Clavien IIIb. Patient Global Impression of Improvement questionnaire scores showed that 6 patients (67%) felt "Very much better" and 3 patients (33%) felt "Much better" after urinary undiversion 60%. CONCLUSION: After urinary undiversions, minor complication rate is high, and major complication rate is considerable. Urinary undiversions are a highly complex, yet feasible procedure, which requires experienced multidisciplinary teams and demands appropriate patient selection. Patients, after undiversions show a high degree of satisfaction with long-term satisfactory outcomes, which points out the need for consideration for these procedures once the oncologic disease is controlled.

Resultados del trasplante renal con donante vivo: análisis comparativo entre arteria renal única y múltiple / Outcome in Living Donor Kidney Transplant: Comparative Análisis of Single and Multiple Renal Arteries

Objetivos: Realizar un análisis comparativo de los resultados funcionales y complicaciones en trasplantados renales y sus respectivos donantes vivos con arteria renal única versus múltiple. Materiales y métodos: Trabajo retrospectivo analítico de una base de datos confeccionada prospectivamente de pacientes sometidos a trasplantes renales con donante vivo en nuestra institución entre mayo de 2010 y julio de 2014. Según el número de arterias presentes en la angiotomografía preoperatoria se confeccionaron dos grupos: grupo 1 (arteria renal única) y grupo 2 (arteria renal múltiple). Resultados: De los 91 pacientes incluidos, el 37% (n=34) presentaba arterias renales múltiples. Se realizó una nefrectomía laparoscópica al 63% (n=36) y el 41% (n=14) de los donantes del grupo 1 y el grupo 2, respectivamente (p=0,05). Tiempos promedio de isquemia total (grupo 1: 55,6 min; grupo 2: 56 min; p=0,931), de anastomosis (grupo 1: 29,6 min; grupo 2: 29,7 min; p=0,982) y de "cirugía de banco" (grupo 1: 23,5 min; grupo 2: 23,8 min; p=0,948). Transfusión de glóbulos rojos en los receptores (grupo 1: 7%; grupo 2: 14%; p=0,23). Porcentaje de hemodiálisis en la primera semana (grupo 1: 5,2%; grupo 2: 5,8%; p=1). No se hallaron diferencias significativas entre grupos a nivel de tasa de complicaciones y días de internación. Ambos grupos presentaron una sobrevida del injerto del 100% a los 35,6 meses de seguimiento promedio. Conclusiones: La presencia de multiplicidad arterial no debe ser considerada una contraindicación para el donante vivo, ya que el implante de estos injertos logra resultados funcionales similares a los injertos renales con arteria única, sin aumentar la morbilidad del procedimiento. (AU)

Objectives: TTo compare functional outcomes and complications in patients with a single artery versus multiple arteries undergoing living donor nephrectomy. Materials and methods: Retrospective analysis of a prospective collected database of living donor kidney transplantations performed at our institution between May 2010 and July 2014. According to the number of arteries present in preoperative angiotomography, two groups of patients were organized: group 1 (single artery) and group 2 (multiple arteries). Results: TNinety-one living donor kidney transplantations were performed during this period. A total of 34 patients (37%) had multiple renal arteries. Mean total ischemia time (single: 55.6 min; multiple: 56 min; p=0.931), anastomosis time (single: 29.6 min; multiple: 29.7 min; p=0.982), bench surgery time (single: 23.5 min; multiple: 23.8 min; p=0.948). Blood transfusion rate was 7% and 14% for group 1 and group 2, respectively (p=0.23). Three patients in group 1 (5.2%) and two patients in group 2 (5.8%) needed dialysis during the first postoperative week (p=1). Overall, recipient complication rate and hospital stay were similar between group 1 and 2. Both groups had a 100% graft survival with a mean follow-up of 35.6 months. Conclusions: The presence of multiple renal arteries should not be considered a contraindication for the living donor nephrectomy, since these grafts achieve similar functional results to single artery renal grafts, without increasing the morbidity of the procedure. (AU)

Resultados oncológicos en pacientes con cáncer de vejiga pT0 posterior a cistectomía radical / Oncological outcomes of Patients with pT0 Bladder Cancer after Radical Cystectomy

Introducción: El estadío pT0 del cáncer de vejiga implica la ausencia de enfermedad en la pieza de cistectomía radical (CR). El objetivo de este estudio es describir los resultados oncológicos de los pacientes con estadío pT0 posterior a CR por carcinoma urotelial de vejiga. Materiales y métodos: Estudio retrospectivo de pacientes sometidos a CR por cáncer de vejiga, en una sola institución, sin neoadyuvancia, entre junio de 2005 y julio de 2013. Se incluyeron aquellos pacientes con diagnóstico histológico de estadío pT0 pN0. Se estimó la sobrevida global, sobrevida cáncer-específica y sobrevida libre de recidiva con el método de Kaplan-Meier. Resultados: De 254 pacientes cistectomizados, 17 presentaron estadío pT0 pN0 (6,7%). La mediana de edad fue 67 años (rango 49-85), 15 pacientes fueron hombres (88%). Los resultados patológicos posterior a resección transuretral de vejiga (RTUv) fueron 17% pT1 (n=3) y 83% pT2 (n=14). La mediana de tiempo entre RTUv y CR fue 60 días (rango 30- 95). Al 41% se le realizó derivación urinaria tipo Bricker y al 59%, una neovejiga. La mediana de estadía hospitalaria fue 8 días (rango 6-44). Se evidenció adenocarcinoma de próstata en 4 pacientes. La mediana de ganglios resecados fue 6 (rango 2-17). Ningún paciente recibió adyuvancia. La mediana de seguimiento fue 69 meses (rango 5-120). Un paciente presentó recidiva uretral a los 72 meses de la CR. La sobrevida cáncer-específica fue 100%, la sobrevida libre de recaída a 5 años fue 83,3% (intervalo de confianza [IC] de 95%: 53,5-100) y la sobrevida global a 5 años fue 82,4% (IC 95%: 64,7-100). Conclusión: El estadío pT0 del cáncer de vejiga presenta resultados oncológicos más favorables que los estadíos más avanzados. Sin embargo, la posibilidad de recurrencia existe, por lo que no se debe discontinuar el seguimiento de estos pacientes (AU)

Introduction: There are cases in which there is no evidence of disease in the radical cystectomy (RC) specimen (pT0 stage). The purpose of this study is to evaluate oncological outcomes of patients with pT0 bladder cancer after RC, in a single institution, without neo-adjuvant therapy. Materials and methods: Patients who underwent radical cystectomy from June 2005 to July 2013 were reviewed retrospectively. All patients had history of bladder urothelial carcinoma, treated with transurethral resection of the bladder (TURB) and confirmed with pathological analysis. Study variables included TURB pathology, time to RC, and pathologic features. Overall survival (OS), cancer-specific survival (CSS) and recurrence-free survival (RFS) were estimated. Results: RC was performed on 254 patients; 17 patients (6.7%) had pT0N0 stage. Median age was 67 years (range 49-85 years); 15 patients were male (88%). TURB pathology specimens were 17% pT1 (n=3), and 83% pT2 (n=14). Median time between TURB and RC was 60 days (range 30-95). Seven patients (41%) received an ileal conduit, and ten patients (59%) received a neobladder. Median hospital stay was eight days (range 6-44). Prostate adenocarcinoma was found in four (23%) patients. Median resected lymph nodes were six (range 2-17). No patients received adjuvant chemotherapy. Median follow-up was 69 months (range 5-120 months). One patient had a urethral relapse 72 months after RC. There was no cancer-specific mortality. RFS at 5 years was 83.3% (confidence interval [CI] 95%: 53.5-100); OS at 5 years was 82.4% (CI 95%: 64.7-100). Conclusion: pT0 stage after radical cystectomy shows more favorable oncologic outcomes than higher stages. However, cancer recurrence was found in a low number of patients, thus, patient follow-up should be maintained (AU)

Preocupación por el bienestar de residentes, un tema presente en la Conferencia Latinoamericana en Educación de Residentes (LACRE) 2017 / Concern for residents' wellbeing, an issue discussed at the latin american conference on resident education (LACRE) 2017

At the end of May 2017, the third version of the Latin American Conference on Resident Education, LACRE, was held in Chile; it convened 433 people from 14 regional countries. Chronic stress and emotional exhaustion of residents was one of the topics discussed. Reports from different countries documented that about half of residents suffer from burnout. This is, they are emotionally drained, indifferent towards their patients and with a sense of low personal fulfillment at work. This article describes the contributions presented in LACRE about interventions or institutional programs designed to reduce burnout and promote self-care of residents. The relevance of these initiatives is discussed in the current global context, considering the available evidence on the effectiveness of interventions to promote well-being among residents. International experts are making renewed and eloquent calls to medical educators and organizations to get involved in the solution of the erosion of resident wellbeing during the residence.

A case of emphysematous cystitis caused by Klebsiella pneumoniae.

Emphysematous cystitis is a rare type of urinary tract infection that is characterized by air pockets within the bladder wall and lumen, which come from gas that is mainly produced by gram-negative bacteria, notably Escherichia coli. This infection is more common in older women with poorly controlled diabetes. An abdominal computerized tomography (CT) scan is the gold standard method to make the diagnosis. The infection can be life-threatening, so prompt treatment is essential. We present a case of a 39-year-old woman with poorly controlled type 2 diabetes who developed emphysematous cystitis after a bilateral adrenalectomy. The infection was diagnosed by a CT scan that revealed gas in the bladder wall. A urine culture revealed 106 colonies/mL of Klebsiella pneumoniae. After a month of treatment with intravenous antibiotics (vancomycin plus meropenem plus colistin), bladder drainage, and strict glycemic control, the patient had a good outcome.

[Concern for residents' wellbeing, an issue discussed at the latin american conference on resident education (LACRE) 2017]. / Preocupación por el bienestar de residentes, un tema presente en la Conferencia Latinoamericana en Educación de Residentes (LACRE) 2017.

At the end of May 2017, the third version of the Latin American Conference on Resident Education, LACRE, was held in Chile; it convened 433 people from 14 regional countries. Chronic stress and emotional exhaustion of residents was one of the topics discussed. Reports from different countries documented that about half of residents suffer from burnout. This is, they are emotionally drained, indifferent towards their patients and with a sense of low personal fulfillment at work. This article describes the contributions presented in LACRE about interventions or institutional programs designed to reduce burnout and promote self-care of residents. The relevance of these initiatives is discussed in the current global context, considering the available evidence on the effectiveness of interventions to promote well-being among residents. International experts are making renewed and eloquent calls to medical educators and organizations to get involved in the solution of the erosion of resident wellbeing during the residence.